Kinetics of the in vitro antibody response to transmissible gastroenteritis (TGE) virus from pig mesenteric lymph node cells, using the ELISASPOT and ELISA tests.
Kinetics of the in vitro antibody response to transmissible gastroenteritis (TGE) virus from pig mesenteric lymph node cells, using the ELISASPOT and ELISA tests.
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DOI：
10.1016/0022-1759(90)90188-2
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发表时间：
1990-08-07
影响因子：
2.2
通讯作者：
Binns RM
中科院分区：
文献类型：
--
作者：
Berthon P;Bernard S;Salmon H;Binns RM
本文描述了猪体内病毒体液免疫反应的体外研究方法。经口服传染性胃肠炎（TGE）冠状病毒免疫后，经病毒抗原体外增强的引物肠系膜淋巴结细胞产生抗体。对于后者，使用多种不同的抗原刺激方法将白细胞与紫外线灭活病毒共培养。采用ELISA法（以3-氨基- 9-乙基咔唑为过氧化物酶显色原）和igg同型分别进行特异性抗体分泌细胞（ASC）计数和抗病毒抗体分泌量测定。结果表明，ASC数与分泌抗体滴度密切相关。在整个培养过程中，致敏细胞与病毒保持接触，体外抗体合成效果最好。抗体反应的动力学特征为窄峰，在培养4天和6天后出现最大值，IgA反应出现的时间早于IgG。这种分析细胞水平特异性抗体反应的方法，可能允许对Ig同型调节机制的研究。扩展到免疫系统其他器官的白细胞，它也可以构成一个体外模型来研究猪不同淋巴组织中表达的抗体反应。
A method is described for in vitro studies of viral humoral immune responses in the pig. After oral immunization with transmissible gastroenteritis (TGE) coronavirus, antibody production from primed mesenteric lymph node cells was revealed by an in vitro boost with viral antigen. For the latter the leukocytes were co-cultured with UV-inactivated virus using a variety of different methods of antigenic stimulation. Enumeration of specific antibody-secreting cells (ASC) and titration of secreted anti-virus antibodies were performed with ELISASPOT (using 3-amino 9-ethyl carbazole as the peroxidase chromogen) and ELISA tests respectively, according to the Ig isotype. The results showed a close relationship between ASC numbers and secreted antibody titres. The best in vitro antibody synthesis was observed when the sensitized cells were maintained in contact with virus during the whole culture period. Antibody responses were defined by a kinetic profile characterized by a narrow peak, with a maximum occuring after 4 and 6 days of culture and with the IgA response appearing earlier than the IgG. This methodology, which analyses specific antibody responses at the cellular level, may permit studies on the mechanisms of Ig isotype regulation. Extended to leukocytes from other organs of the immune system, it may also constitute an in vitro model to study antibody responses expressed in different lymphoid tissues of the pig.
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影响因子：
2.2
作者：
CZERKINSKY, CC;NILSSON, LA;TARKOWSKI, A
通讯作者：
TARKOWSKI, A
DOI：
10.1016/s0769-2617(87)80019-9
发表时间：
1987-07-01
期刊：
ANNALES DE L INSTITUT PASTEUR-VIROLOGY
影响因子：
--
作者：
NGUYEN, TD;BOTTREAU, E;AYNAUD, JM
通讯作者：
AYNAUD, JM
影响因子：
3.8
作者：
AYNAUD, JM;NGUYEN, TD;VANNIER, P
通讯作者：
VANNIER, P
影响因子：
2.2
作者：
BERNARD, S;LANTIER, I
通讯作者：
LANTIER, I
DOI：
10.1016/s0769-2617(87)80018-7
发表时间：
1987-07-01
期刊：
ANNALES DE L INSTITUT PASTEUR-VIROLOGY
影响因子：
--
作者：
NGUYEN, TD;BERNARD, S;AYNAUD, JM
通讯作者：
AYNAUD, JM